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Most alphaviruses are mosquito-borne and can cause severe disease in humans and domesticated animals. In North America, eastern equine encephalitis virus (EEEV) is an important human pathogen with case fatality rates of 30–90%. Currently, there are no therapeutics or vaccines to treat and/or prevent human infection. One critical impediment in countermeasure development is the lack of insight into clinically relevant parameters in a susceptible animal model. This study examined the disease course of EEEV in a cynomolgus macaque model utilizing advanced telemetry technology to continuously and simultaneously measure temperature, respiration, activity, heart rate, blood pressure, electrocardiogram (ECG), and electroencephalography (EEG) following an aerosol challenge at 7.0 log10 PFU. Following challenge, all parameters were rapidly and substantially altered with peak alterations from baseline ranged as follows: temperature (+3.0–4.2°C), respiration rate (+56–128%), activity (-15-76% daytime and +5–22% nighttime), heart rate (+67–190%), systolic (+44–67%) and diastolic blood pressure (+45–80%). Cardiac abnormalities comprised of alterations in QRS and PR duration, QTc Bazett, T wave morphology, amplitude of the QRS complex, and sinoatrial arrest. An unexpected finding of the study was the first documented evidence of a critical cardiac event as an immediate cause of euthanasia in one NHP. All brain waves were rapidly (~12–24 hpi) and profoundly altered with increases of up to 6,800% and severe diffuse slowing of all waves with decreases of ~99%. Lastly, all NHPs exhibited disruption of the circadian rhythm, sleep, and food/fluid intake. Accordingly, all NHPs met the euthanasia criteria by ~106–140 hpi. This is the first of its kind study utilizing state of the art telemetry to investigate multiple clinical parameters relevant to human EEEV infection in a susceptible cynomolgus macaque model. The study provides critical insights into EEEV pathogenesis and the parameters identified will improve animal model development to facilitate rapid evaluation of vaccines and therapeutics.
John C. Trefry; Franco D. Rossi; Michael V. Accardi; Brandi L. Dorsey; Thomas R. Sprague; Suzanne E. Wollen-Roberts; Joshua D. Shamblin; Adrienne E. Kimmel; Pamela J. Glass; Lynn J. Miller; Crystal W. Burke; Anthony P. Cardile; Darci R. Smith; Sina Bavari; Simon Authier; William D. Pratt; Margaret L. Pitt; Farooq Nasar. The utilization of advance telemetry to investigate critical physiological parameters including electroencephalography in cynomolgus macaques following aerosol challenge with eastern equine encephalitis virus. PLOS Neglected Tropical Diseases 2021, 15, e0009424 .
AMA StyleJohn C. Trefry, Franco D. Rossi, Michael V. Accardi, Brandi L. Dorsey, Thomas R. Sprague, Suzanne E. Wollen-Roberts, Joshua D. Shamblin, Adrienne E. Kimmel, Pamela J. Glass, Lynn J. Miller, Crystal W. Burke, Anthony P. Cardile, Darci R. Smith, Sina Bavari, Simon Authier, William D. Pratt, Margaret L. Pitt, Farooq Nasar. The utilization of advance telemetry to investigate critical physiological parameters including electroencephalography in cynomolgus macaques following aerosol challenge with eastern equine encephalitis virus. PLOS Neglected Tropical Diseases. 2021; 15 (6):e0009424.
Chicago/Turabian StyleJohn C. Trefry; Franco D. Rossi; Michael V. Accardi; Brandi L. Dorsey; Thomas R. Sprague; Suzanne E. Wollen-Roberts; Joshua D. Shamblin; Adrienne E. Kimmel; Pamela J. Glass; Lynn J. Miller; Crystal W. Burke; Anthony P. Cardile; Darci R. Smith; Sina Bavari; Simon Authier; William D. Pratt; Margaret L. Pitt; Farooq Nasar. 2021. "The utilization of advance telemetry to investigate critical physiological parameters including electroencephalography in cynomolgus macaques following aerosol challenge with eastern equine encephalitis virus." PLOS Neglected Tropical Diseases 15, no. 6: e0009424.
Most alphaviruses are mosquito-borne and can cause severe disease in humans and domesticated animals. In North America, eastern equine encephalitis virus (EEEV) is an important human pathogen with case fatality rates of 30-90%. Currently, there are no therapeutics or vaccines to treat and/or prevent human infection. One critical impediment in countermeasure development is the lack of insight into clinically relevant parameters in a susceptible animal model. This study examined the disease course of EEEV in a cynomolgus macaque model utilizing advanced telemetry technology to continuously and simultaneously measure temperature, respiration, activity, heart rate, blood pressure, electrocardiogram (ECG), and electroencephalography (EEG) following an aerosol challenge at 7.0 log10 PFU. Following challenge, all parameters were rapidly and substantially altered with peak alterations from baseline ranged as follows: temperature (+3.0-4.2 °C), respiration rate (+56-128%), activity (+15-76% daytime and +5-22% nighttime), heart rate (+67-190%), systolic (+44-67%) and diastolic blood pressure (+45-80%). Cardiac abnormalities comprised of alterations in QRS and PR duration, QTc Bazett, T wave morphology, amplitude of the QRS complex, and sinoatrial arrest. An unexpected finding of the study was the first documented evidence of a critical cardiac event as an immediate cause of euthanasia in one NHP. All brain waves were rapidly (12-24 hpi) and profoundly altered with increases of up to 6,800% and severe diffuse slowing of all waves with decreases of ∼99%. Lastly, all NHPs exhibited disruption of the circadian rhythm, sleep, and food/fluid intake. Accordingly, all NHPs met the euthanasia criteria by ∼106-140 hpi. This is the first of its kind study utilizing state of the art telemetry to investigate multiple clinical parameters relevant to human EEEV infection in a susceptible cynomolgus macaque model. The study provides critical insights into EEEV pathogenesis and the parameters identified will improve animal model development to facilitate rapid evaluation of vaccines and therapeutics.Author SummaryIn North America, EEEV causes the most severe mosquito-borne disease in humans highlighted by fatal encephalitis and permeant debilitating neurological sequelae in survivors. The first confirmed human cases were reported more than 80 years ago and since then multiple sporadic outbreaks have occurred including one of the largest in 2019. Unfortunately, most human infections are diagnosed at the on-set of severe neurological symptoms and consequently a detailed disease course in humans is lacking. This gap in knowledge is a significant obstacle in the development of appropriate animal models to evaluate countermeasures. Here, we performed a cutting-edge study by utilizing a new telemetry technology to understand the course of EEEV infection in a susceptible macaque model by measuring multiple physiological parameters relevant to human disease. Our study demonstrates that the infection rapidly produces considerable alterations in many critical parameters including the electrical activity of the heart and the brain leading to severe disease. The study also highlights the extraordinary potential of new telemetry technology to develop the next generation of animal models in order to comprehensively investigate pathogenesis as well as evaluate countermeasures to treat and/or prevent EEEV disease.
John C Trefry; Franco D Rossi; Michael V Accardi; Brandi L Dorsey; Thomas R Sprague; Suzanne E Wollen-Roberts; Joshua D Shamblin; Adrienne E Kimmel; Pamela J Glass; Lynn J Miller; Anthony P Cardile; Darci R Smith; Sina Bavari; Simon Authier; William D Pratt; Margaret L Pitt; Farooq Nasar. The Utilization of Advance Telemetry to Investigate Important Physiological Parameters Including Electroencephalography in Cynomolgus Macaques Following Aerosol Challenge with Eastern Equine Encephalitis Virus. 2020, 1 .
AMA StyleJohn C Trefry, Franco D Rossi, Michael V Accardi, Brandi L Dorsey, Thomas R Sprague, Suzanne E Wollen-Roberts, Joshua D Shamblin, Adrienne E Kimmel, Pamela J Glass, Lynn J Miller, Anthony P Cardile, Darci R Smith, Sina Bavari, Simon Authier, William D Pratt, Margaret L Pitt, Farooq Nasar. The Utilization of Advance Telemetry to Investigate Important Physiological Parameters Including Electroencephalography in Cynomolgus Macaques Following Aerosol Challenge with Eastern Equine Encephalitis Virus. . 2020; ():1.
Chicago/Turabian StyleJohn C Trefry; Franco D Rossi; Michael V Accardi; Brandi L Dorsey; Thomas R Sprague; Suzanne E Wollen-Roberts; Joshua D Shamblin; Adrienne E Kimmel; Pamela J Glass; Lynn J Miller; Anthony P Cardile; Darci R Smith; Sina Bavari; Simon Authier; William D Pratt; Margaret L Pitt; Farooq Nasar. 2020. "The Utilization of Advance Telemetry to Investigate Important Physiological Parameters Including Electroencephalography in Cynomolgus Macaques Following Aerosol Challenge with Eastern Equine Encephalitis Virus." , no. : 1.
Eastern equine encephalitis virus (EEEV) is mosquito-borne virus that produces fatal encephalitis in humans. We recently conducted a first of its kind study to investigate EEEV clinical disease course following aerosol challenge in a cynomolgus macaque model utilizing the state of the art telemetry to measure critical physiological parameters. Following challenge, all parameters were rapidly and profoundly altered, and all nonhuman primates (NHPs) met the euthanasia criteria. In this study, we performed the first comprehensive pathology investigation of tissues collected at euthanasia to gain insights into EEEV pathogenesis. Viral RNA and proteins as well as microscopic lesions were absent in the visceral organs. In contrast, viral RNA and proteins were readily detected throughout the brain including autonomic nervous system (ANS) control centers and spinal cord. However, despite presence of viral RNA and proteins, majority of the brain and spinal cord tissues exhibited minimal or no microscopic lesions. The virus tropism was restricted primarily to neurons, and virus particles (~61-68 nm) were present within axons of neurons and throughout the extracellular spaces. However, active virus replication was absent or minimal in majority of the brain and was limited to regions proximal to the olfactory tract. These data suggest that EEEV initially replicates in/near the olfactory bulb following aerosol challenge and is rapidly transported to distal regions of the brain by exploiting the neuronal axonal transport system to facilitate neuron-to-neuron spread. Once within the brain, the virus gains access to the ANS control centers likely leading to disruption and/or dysregulation of critical physiological parameters to produce severe disease. Moreover, the absence of microscopic lesions strongly suggests that the underlying mechanism of EEEV pathogenesis is due to neuronal dysfunction rather than neuronal death. This study is the first comprehensive investigation of EEEV clinical disease course and pathogenesis in a NHP model and will provide significant insights into the evaluation of countermeasure.
Janice A Williams; Simon Y Long; Xiankun Zeng; Kathleen Kuehl; April M Babka; Neil M Davis; Jun Liu; John C Trefry; Sharon Daye; Paul R Facemire; Patrick L Iversen; Sina Bavari; Margaret L Pitt; Farooq Nasar. Eastern Equine Encephalitis Virus Rapidly Infects and Disseminates in the Brain and Spinal Cord of Infected Cynomolgus Macaques Following Aerosol Challenge. 2020, 1 .
AMA StyleJanice A Williams, Simon Y Long, Xiankun Zeng, Kathleen Kuehl, April M Babka, Neil M Davis, Jun Liu, John C Trefry, Sharon Daye, Paul R Facemire, Patrick L Iversen, Sina Bavari, Margaret L Pitt, Farooq Nasar. Eastern Equine Encephalitis Virus Rapidly Infects and Disseminates in the Brain and Spinal Cord of Infected Cynomolgus Macaques Following Aerosol Challenge. . 2020; ():1.
Chicago/Turabian StyleJanice A Williams; Simon Y Long; Xiankun Zeng; Kathleen Kuehl; April M Babka; Neil M Davis; Jun Liu; John C Trefry; Sharon Daye; Paul R Facemire; Patrick L Iversen; Sina Bavari; Margaret L Pitt; Farooq Nasar. 2020. "Eastern Equine Encephalitis Virus Rapidly Infects and Disseminates in the Brain and Spinal Cord of Infected Cynomolgus Macaques Following Aerosol Challenge." , no. : 1.
Venezuelan equine encephalitis virus (VEEV) is an important pathogen of medical and veterinary importance in the Americas. In this report, we present the complete genome sequences of five VEEV isolates obtained from pools of Culex (Melanoconion) gnomatos (4) or Culex (Melanoconion) pedroi (1) from Iquitos, Peru. Genetic and phylogenetic analyses showed that all five isolates grouped within the VEEV complex sister to VEEV IIIC and are members of subtype IIID. This is the first report of full-length genomic sequences of VEEV IIID.
Susana L. Padilla; Karla Prieto; David J. Dohm; Michael J. Turell; Terry A. Klein; Roberto Fernández; Douglas M. Watts; Robert G. Lowen; Gustavo F. Palacios; Margaret L. Pitt; Michael R. Wiley; Farooq Nasar. Complete genomic sequences of Venezuelan equine encephalitis virus subtype IIID isolates from mosquitoes. Archives of Virology 2020, 165, 1715 -1717.
AMA StyleSusana L. Padilla, Karla Prieto, David J. Dohm, Michael J. Turell, Terry A. Klein, Roberto Fernández, Douglas M. Watts, Robert G. Lowen, Gustavo F. Palacios, Margaret L. Pitt, Michael R. Wiley, Farooq Nasar. Complete genomic sequences of Venezuelan equine encephalitis virus subtype IIID isolates from mosquitoes. Archives of Virology. 2020; 165 (7):1715-1717.
Chicago/Turabian StyleSusana L. Padilla; Karla Prieto; David J. Dohm; Michael J. Turell; Terry A. Klein; Roberto Fernández; Douglas M. Watts; Robert G. Lowen; Gustavo F. Palacios; Margaret L. Pitt; Michael R. Wiley; Farooq Nasar. 2020. "Complete genomic sequences of Venezuelan equine encephalitis virus subtype IIID isolates from mosquitoes." Archives of Virology 165, no. 7: 1715-1717.
Licensure of a vaccine to protect against aerosolized Venezuelan equine encephalitis virus (VEEV) requires use of the U.S. Food and Drug Administration (FDA) Animal Rule to assess vaccine efficacy as human studies are not feasible or ethical. An approach to selecting VEEV challenge strains for use under the Animal Rule was developed, taking into account Department of Defense (DOD) vaccine requirements, FDA Animal Rule guidelines, strain availability, and lessons learned from the generation of filovirus challenge agents within the Filovirus Animal Nonclinical Group (FANG). Initial down-selection to VEEV IAB and IC epizootic varieties was based on the DOD objective for vaccine protection in a bioterrorism event. The subsequent down-selection of VEEV IAB and IC isolates was based on isolate availability, origin, virulence, culture and animal passage history, known disease progression in animal models, relevancy to human disease, and ability to generate sufficient challenge material. Methods for the propagation of viral stocks (use of uncloned (wild-type), plaque-cloned, versus cDNA-cloned virus) to minimize variability in the potency of the resulting challenge materials were also reviewed. The presented processes for VEEV strain selection and the propagation of viral stocks may serve as a template for animal model development product testing under the Animal Rule to other viral vaccine programs. This manuscript is based on the culmination of work presented at the “Alphavirus Workshop” organized and hosted by the Joint Vaccine Acquisition Program (JVAP) on 15 December 2014 at Fort Detrick, Maryland, USA.
Janice M. Rusnak; Pamela J. Glass; Scott C. Weaver; Carol L. Sabourin; Andrew M. Glenn; William Klimstra; Christopher S. Badorrek; Farooq Nasar; Lucy A. Ward. Approach to Strain Selection and the Propagation of Viral Stocks for Venezuelan Equine Encephalitis Virus Vaccine Efficacy Testing under the Animal Rule. Viruses 2019, 11, 807 .
AMA StyleJanice M. Rusnak, Pamela J. Glass, Scott C. Weaver, Carol L. Sabourin, Andrew M. Glenn, William Klimstra, Christopher S. Badorrek, Farooq Nasar, Lucy A. Ward. Approach to Strain Selection and the Propagation of Viral Stocks for Venezuelan Equine Encephalitis Virus Vaccine Efficacy Testing under the Animal Rule. Viruses. 2019; 11 (9):807.
Chicago/Turabian StyleJanice M. Rusnak; Pamela J. Glass; Scott C. Weaver; Carol L. Sabourin; Andrew M. Glenn; William Klimstra; Christopher S. Badorrek; Farooq Nasar; Lucy A. Ward. 2019. "Approach to Strain Selection and the Propagation of Viral Stocks for Venezuelan Equine Encephalitis Virus Vaccine Efficacy Testing under the Animal Rule." Viruses 11, no. 9: 807.
Recent advances in mass spectrometry methods and instrumentation now allow for more accurate identification of proteins in low abundance. This technology was applied to Sindbis virus, the prototypical alphavirus, to investigate the viral proteome. To determine if host proteins are specifically packaged into alphavirus virions, Sindbis virus (SINV) was grown in multiple host cells representing vertebrate and mosquito hosts, and total protein content of purified virions was determined. This analysis identified host factors not previously associated with alphavirus entry, replication, or egress. One host protein, sorting nexin 5 (SNX5), was shown to be critical for the replication of three different alphaviruses, Sindbis, Mayaro, and Chikungunya viruses. The most significant finding was that in addition to the host proteins, SINV nonstructural protein 2 (nsP2) was detected within virions grown in all host cells examined. The protein and RNA-interacting capabilities of nsP2 coupled with its presence in the virion support a role for nsP2 during packaging and/or entry of progeny virus. This function has not been identified for this protein. Taken together, this strategy identified at least one host factor integrally involved in alphavirus replication. Identification of other host proteins provides insight into alphavirus-host interactions during viral replication in both vertebrate and invertebrate hosts. This method of virus proteome analysis may also be useful for the identification of protein candidates for host-based therapeutics. IMPORTANCE Pathogenic alphaviruses, such as Chikungunya and Mayaro viruses, continue to plague public health in developing and developed countries alike. Alphaviruses belong to a group of viruses vectored in nature by hematophagous (blood-feeding) insects and are termed arboviruses (arthropod-borne viruses). This group of viruses contains many human pathogens, such as dengue fever, West Nile, and Yellow fever viruses. With few exceptions, there are no vaccines or prophylactics for these agents, leaving one-third of the world population at risk of infection. Identifying effective antivirals has been a long-term goal for combating these diseases not only because of the lack of vaccines but also because they are effective during an ongoing epidemic. Mass spectrometry-based analysis of the Sindbis virus proteome can be effective in identifying host genes involved in virus replication and novel functions for virus proteins. Identification of these factors is invaluable for the prophylaxis of this group of viruses.
Ryan Schuchman; Andy Kilianski; Amanda Piper; Ricardo Vancini; José M. C. Ribeiro; Thomas R. Sprague; Farooq Nasar; Gabrielle Boyd; Raquel Hernandez; Trevor Glaros. Comparative Characterization of the Sindbis Virus Proteome from Mammalian and Invertebrate Hosts Identifies nsP2 as a Component of the Virion and Sorting Nexin 5 as a Significant Host Factor for Alphavirus Replication. Journal of Virology 2018, 92, 1 .
AMA StyleRyan Schuchman, Andy Kilianski, Amanda Piper, Ricardo Vancini, José M. C. Ribeiro, Thomas R. Sprague, Farooq Nasar, Gabrielle Boyd, Raquel Hernandez, Trevor Glaros. Comparative Characterization of the Sindbis Virus Proteome from Mammalian and Invertebrate Hosts Identifies nsP2 as a Component of the Virion and Sorting Nexin 5 as a Significant Host Factor for Alphavirus Replication. Journal of Virology. 2018; 92 (14):1.
Chicago/Turabian StyleRyan Schuchman; Andy Kilianski; Amanda Piper; Ricardo Vancini; José M. C. Ribeiro; Thomas R. Sprague; Farooq Nasar; Gabrielle Boyd; Raquel Hernandez; Trevor Glaros. 2018. "Comparative Characterization of the Sindbis Virus Proteome from Mammalian and Invertebrate Hosts Identifies nsP2 as a Component of the Virion and Sorting Nexin 5 as a Significant Host Factor for Alphavirus Replication." Journal of Virology 92, no. 14: 1.
The Togaviridae is a family of small, enveloped viruses with single-stranded, positive-sense RNA genomes of 10–12 kb. Within the family, the genus Alphavirus includes a large number of diverse species, while the genus Rubivirus includes the single species Rubella virus. Most alphaviruses are mosquito-borne and are pathogenic in their vertebrate hosts. Many are important human and veterinary pathogens (e.g. chikungunya virus and eastern equine encephalitis virus). Rubella virus is transmitted by respiratory routes among humans. This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the taxonomy of the Togaviridae, which is available at www.ictv.global/report/togaviridae.
Rubing Chen; Suchetana Mukhopadhyay; Andres Merits; Bethany Bolling; Farooq Nasar; Lark L. Coffey; Ann Powers; Scott Weaver; ICTV Report Consortium. ICTV Virus Taxonomy Profile: Togaviridae. Journal of General Virology 2018, 99, 761 -762.
AMA StyleRubing Chen, Suchetana Mukhopadhyay, Andres Merits, Bethany Bolling, Farooq Nasar, Lark L. Coffey, Ann Powers, Scott Weaver, ICTV Report Consortium. ICTV Virus Taxonomy Profile: Togaviridae. Journal of General Virology. 2018; 99 (6):761-762.
Chicago/Turabian StyleRubing Chen; Suchetana Mukhopadhyay; Andres Merits; Bethany Bolling; Farooq Nasar; Lark L. Coffey; Ann Powers; Scott Weaver; ICTV Report Consortium. 2018. "ICTV Virus Taxonomy Profile: Togaviridae." Journal of General Virology 99, no. 6: 761-762.
The United States Army Medical Research Institute of Infectious Diseases (USAMRIID) possesses an array of expertise in diverse capabilities for the characterization of emerging infectious diseases from the pathogen itself to human or animal infection models. The recent Zika virus (ZIKV) outbreak was a challenge and an opportunity to put these capabilities to work as a cohesive unit to quickly respond to a rapidly developing threat. Next-generation sequencing was used to characterize virus stocks and to understand the introduction and spread of ZIKV in the United States. High Content Imaging was used to establish a High Content Screening process to evaluate antiviral therapies. Functional genomics was used to identify critical host factors for ZIKV infection. An animal model using the temporal blockade of IFN-I in immunocompetent laboratory mice was investigated in conjunction with Positron Emission Tomography to study ZIKV. Correlative light and electron microscopy was used to examine ZIKV interaction with host cells in culture and infected animals. A quantitative mass spectrometry approach was used to examine the protein and metabolite type or concentration changes that occur during ZIKV infection in blood, cells, and tissues. Multiplex fluorescence in situ hybridization was used to confirm ZIKV replication in mouse and NHP tissues. The integrated rapid response approach developed at USAMRIID presented in this review was successfully applied and provides a new template pathway to follow if a new biological threat emerges. This streamlined approach will increase the likelihood that novel medical countermeasures could be rapidly developed, evaluated, and translated into the clinic.
Robert G. Lowen; Thomas M. Bocan; Christopher Kane; Lisa Cazares; Krishna P. Kota; Jason T. Ladner; Farooq Nasar; Margaret Louise Pitt; Darci R. Smith; Veronica Soloveva; Mei G. Sun; Xiankun (Kevin) Zeng; Sina Bavari. Countering Zika Virus: The USAMRIID Response. Advances in Experimental Medicine and Biology 2018, 1062, 303 -318.
AMA StyleRobert G. Lowen, Thomas M. Bocan, Christopher Kane, Lisa Cazares, Krishna P. Kota, Jason T. Ladner, Farooq Nasar, Margaret Louise Pitt, Darci R. Smith, Veronica Soloveva, Mei G. Sun, Xiankun (Kevin) Zeng, Sina Bavari. Countering Zika Virus: The USAMRIID Response. Advances in Experimental Medicine and Biology. 2018; 1062 ():303-318.
Chicago/Turabian StyleRobert G. Lowen; Thomas M. Bocan; Christopher Kane; Lisa Cazares; Krishna P. Kota; Jason T. Ladner; Farooq Nasar; Margaret Louise Pitt; Darci R. Smith; Veronica Soloveva; Mei G. Sun; Xiankun (Kevin) Zeng; Sina Bavari. 2018. "Countering Zika Virus: The USAMRIID Response." Advances in Experimental Medicine and Biology 1062, no. : 303-318.
To evaluate potential immunocompetent small animal models of Zika virus (ZIKV) infection, we inoculated Syrian golden hamsters (subcutaneously or intraperitoneally) and strain 13 guinea pigs (intraperitoneally) with Senegalese ZIKV strain ArD 41525 or Philippines ZIKV strain CPC-0740. We did not detect viremia in hamsters inoculated subcutaneously with either virus strain, although some hamsters developed virus neutralizing antibodies. However, we detected statistically significant higher viremias (P = 0.0285) and a higher median neutralization titer (P = 0.0163) in hamsters inoculated intraperitoneally with strain ArD 41525 compared with strain CPC-0740. Furthermore, some hamsters inoculated with strain ArD 41525 displayed mild signs of disease. By contrast, strain 13 guinea pigs inoculated intraperitoneally with either strain did not have detectable viremias and less than half developed virus neutralizing antibodies. Our results support the use of the Syrian golden hamster intraperitoneal model to explore phenotypic variation between ZIKV strains.
Lynn J. Miller; Farooq Nasar; Christopher W. Schellhase; Sarah L. Norris; Adrienne Kimmel; Stephanie M. Valdez; Suzanne E. Wollen-Roberts; Joshua D. Shamblin; Thomas R. Sprague; Luis A. Lugo-Roman; Richard G. Jarman; In-Kyu Yoon; Maria T. Alera; Sina Bavari; Margaret Louise Pitt; Andrew D. Haddow. Zika Virus Infection in Syrian Golden Hamsters and Strain 13 Guinea Pigs. The American Journal of Tropical Medicine and Hygiene 2018, 98, 864 -867.
AMA StyleLynn J. Miller, Farooq Nasar, Christopher W. Schellhase, Sarah L. Norris, Adrienne Kimmel, Stephanie M. Valdez, Suzanne E. Wollen-Roberts, Joshua D. Shamblin, Thomas R. Sprague, Luis A. Lugo-Roman, Richard G. Jarman, In-Kyu Yoon, Maria T. Alera, Sina Bavari, Margaret Louise Pitt, Andrew D. Haddow. Zika Virus Infection in Syrian Golden Hamsters and Strain 13 Guinea Pigs. The American Journal of Tropical Medicine and Hygiene. 2018; 98 (3):864-867.
Chicago/Turabian StyleLynn J. Miller; Farooq Nasar; Christopher W. Schellhase; Sarah L. Norris; Adrienne Kimmel; Stephanie M. Valdez; Suzanne E. Wollen-Roberts; Joshua D. Shamblin; Thomas R. Sprague; Luis A. Lugo-Roman; Richard G. Jarman; In-Kyu Yoon; Maria T. Alera; Sina Bavari; Margaret Louise Pitt; Andrew D. Haddow. 2018. "Zika Virus Infection in Syrian Golden Hamsters and Strain 13 Guinea Pigs." The American Journal of Tropical Medicine and Hygiene 98, no. 3: 864-867.
Most alphaviruses are mosquito borne and exhibit a broad host range, infecting many different vertebrates, including birds, rodents, equids, humans, and nonhuman primates. Recently, a host-restricted, mosquito-borne alphavirus, Eilat virus (EILV), was described with an inability to infect vertebrate cells based on defective attachment and/or entry, as well as a lack of genomic RNA replication. We investigated the utilization of EILV recombinant technology as a vaccine platform against eastern (EEEV) and Venezuelan equine encephalitis viruses (VEEV), two important pathogens of humans and domesticated animals. EILV chimeras containing structural proteins of EEEV or VEEV were engineered and successfully rescued in Aedes albopictus cells. Cryo-electron microscopy reconstructions at 8 and 11 Å of EILV/VEEV and EILV/EEEV, respectively, showed virion and glycoprotein spike structures similar to those of VEEV-TC83 and other alphaviruses. The chimeras were unable to replicate in vertebrate cell lines or in brains of newborn mice when injected intracranially. Histopathologic examinations of the brain tissues showed no evidence of pathological lesions and were indistinguishable from those of mock-infected animals. A single-dose immunization of either monovalent or multivalent EILV chimera(s) generated neutralizing antibody responses and protected animals against lethal challenge 70 days later. Lastly, a single dose of monovalent EILV chimeras generated protective responses as early as day 1 postvaccination and partial or complete protection by day 6. These data demonstrate the safety, immunogenicity, and efficacy of novel insect-specific EILV-based chimeras as potential EEEV and VEEV vaccines. IMPORTANCE Mostly in the last decade, insect-specific viruses have been discovered in several arbovirus families. However, most of these viruses are not well studied and largely have been ignored. We explored the use of the mosquito-specific alphavirus EILV as an alphavirus vaccine platform in well-established disease models for eastern (EEE) and Venezuelan equine encephalitis (VEE). EILV-based chimeras replicated to high titers in a mosquito cell line yet retained their host range restriction in vertebrates both in vitro and in vivo . In addition, the chimeras generated immune responses that were higher than those of other human and/or equine vaccines. These findings indicate the feasibility of producing a safe, efficacious, mono- or multivalent vaccine against the encephalitic alphaviruses VEEV and EEEV. Lastly, these data demonstrate how host-restricted, insect-specific viruses can be engineered to develop vaccines against related pathogenic arboviruses that cause severe disease in humans and domesticated animals.
Jesse H. Erasmus; Robert L. Seymour; Jason T. Kaelber; Dal Y. Kim; Grace Leal; Michael B. Sherman; Ilya Frolov; Wah Chiu; Scott C. Weaver; Farooq Nasar. Novel Insect-Specific Eilat Virus-Based Chimeric Vaccine Candidates Provide Durable, Mono- and Multivalent, Single-Dose Protection against Lethal Alphavirus Challenge. Journal of Virology 2018, 92, e01274-17 .
AMA StyleJesse H. Erasmus, Robert L. Seymour, Jason T. Kaelber, Dal Y. Kim, Grace Leal, Michael B. Sherman, Ilya Frolov, Wah Chiu, Scott C. Weaver, Farooq Nasar. Novel Insect-Specific Eilat Virus-Based Chimeric Vaccine Candidates Provide Durable, Mono- and Multivalent, Single-Dose Protection against Lethal Alphavirus Challenge. Journal of Virology. 2018; 92 (4):e01274-17.
Chicago/Turabian StyleJesse H. Erasmus; Robert L. Seymour; Jason T. Kaelber; Dal Y. Kim; Grace Leal; Michael B. Sherman; Ilya Frolov; Wah Chiu; Scott C. Weaver; Farooq Nasar. 2018. "Novel Insect-Specific Eilat Virus-Based Chimeric Vaccine Candidates Provide Durable, Mono- and Multivalent, Single-Dose Protection against Lethal Alphavirus Challenge." Journal of Virology 92, no. 4: e01274-17.
Zika virus (ZIKV) is a mosquito-borne member of the genus Flavivirus that has emerged since 2007 to cause outbreaks in Africa, Asia, Oceania, and most recently, in the Americas. Here, we used an isolate history as well as genetic and phylogenetic analyses to characterize three low-passage isolates representing African (ArD 41525) and Asian (CPC-0740, SV0127-14) lineages to investigate the potential phenotypic differences in vitro and in vivo. The African isolate displayed a large plaque phenotype (∼3–4 mm) on Vero and HEK-293 cells, whereas the Asian isolates either exhibited a small plaque phenotype (∼1–2 mm) or did not produce any plaques. In multistep replication kinetics in nine different vertebrate and insect cell lines, the African isolate consistently displayed faster replication kinetics and yielded ∼10- to 10,000-fold higher peak virus titers (infectious or RNA copies) compared with the Asian isolates. Oral exposure of Aedes aegypti mosquitoes with the African isolate yielded higher infection and dissemination rates compared with the Asian isolates. Infection of Ifnar1 −/− mice with the African isolate produced a uniformly fatal disease, whereas infection with the Asian isolates produced either a delay in time-to-death or a significantly lower mortality rate. Last, the African isolate was > 10,000-fold more virulent than the Asian isolates in an interferon type I antibody blockade mouse model. These data demonstrate substantial phenotypic differences between low-passage African and Asian isolates both in vitro and in vivo and warrant further investigation. They also highlight the need for basic characterization of ZIKV isolates, as the utilization of the uncharacterized isolates could have consequences for animal model and therapeutic/vaccine development.
Darci R. Smith; David A. Kulesh; Stephanie A. Bellanca; Andrew D. Haddow; Thomas R. Sprague; Rome Buathong; Timothy D. Minogue; Lynn Jean Miller; Maria Theresa P. Alera; Stephanie M. Valdez; Farooq Nasar; Susan R. Coyne; Margaret L. Pitt; Richard G. Jarman; Kenneth J. Linthicum; Scott C. Weaver; Joseph W. Golden; Jeff W. Koehler; Bradley S. Hollidge; Gregory D. Gromowski; Christopher D. Kane; Robert B. Tesh; Robert G. Lowen; In-Kyu Yoon; Sina Bavari; Susana L. Padilla. African and Asian Zika Virus Isolates Display Phenotypic Differences Both In Vitro and In Vivo. The American Journal of Tropical Medicine and Hygiene 2018, 98, 432 -444.
AMA StyleDarci R. Smith, David A. Kulesh, Stephanie A. Bellanca, Andrew D. Haddow, Thomas R. Sprague, Rome Buathong, Timothy D. Minogue, Lynn Jean Miller, Maria Theresa P. Alera, Stephanie M. Valdez, Farooq Nasar, Susan R. Coyne, Margaret L. Pitt, Richard G. Jarman, Kenneth J. Linthicum, Scott C. Weaver, Joseph W. Golden, Jeff W. Koehler, Bradley S. Hollidge, Gregory D. Gromowski, Christopher D. Kane, Robert B. Tesh, Robert G. Lowen, In-Kyu Yoon, Sina Bavari, Susana L. Padilla. African and Asian Zika Virus Isolates Display Phenotypic Differences Both In Vitro and In Vivo. The American Journal of Tropical Medicine and Hygiene. 2018; 98 (2):432-444.
Chicago/Turabian StyleDarci R. Smith; David A. Kulesh; Stephanie A. Bellanca; Andrew D. Haddow; Thomas R. Sprague; Rome Buathong; Timothy D. Minogue; Lynn Jean Miller; Maria Theresa P. Alera; Stephanie M. Valdez; Farooq Nasar; Susan R. Coyne; Margaret L. Pitt; Richard G. Jarman; Kenneth J. Linthicum; Scott C. Weaver; Joseph W. Golden; Jeff W. Koehler; Bradley S. Hollidge; Gregory D. Gromowski; Christopher D. Kane; Robert B. Tesh; Robert G. Lowen; In-Kyu Yoon; Sina Bavari; Susana L. Padilla. 2018. "African and Asian Zika Virus Isolates Display Phenotypic Differences Both In Vitro and In Vivo." The American Journal of Tropical Medicine and Hygiene 98, no. 2: 432-444.
There is an urgent need for therapeutic development to combat infections caused by Rift Valley fever virus (RVFV), which causes devastating disease in both humans and animals. In an effort to repurpose drugs for RVFV treatment, our previous studies screened a library of FDA-approved drugs. The most promising candidate identified was the hepatocellular and renal cell carcinoma drug sorafenib. Mechanism-of-action studies indicated that sorafenib targeted a late stage in virus infection and caused a buildup of virions within cells. In addition, small interfering RNA (siRNA) knockdown studies suggested that nonclassical targets of sorafenib are important for the propagation of RVFV. Here we extend our previous findings to identify the mechanism by which sorafenib inhibits the release of RVFV virions from the cell. Confocal microscopy imaging revealed that glycoprotein Gn colocalizes and accumulates within the endoplasmic reticulum (ER) and the transport of Gn from the Golgi complex to the host cell membrane is reduced. Transmission electron microscopy demonstrated that sorafenib caused virions to be present inside large vacuoles inside the cells. p97/valosin-containing protein (VCP), which is involved in membrane remodeling in the secretory pathway and a known target of sorafenib, was found to be important for RVFV egress. Knockdown of VCP resulted in decreased RVFV replication, reduced Gn Golgi complex localization, and increased Gn ER accumulation. The intracellular accumulation of RVFV virions was also observed in cells transfected with siRNA targeting VCP. Collectively, these data indicate that sorafenib causes a disruption in viral egress by targeting VCP and the secretory pathway, resulting in a buildup of virions within dilated ER vesicles. IMPORTANCE In humans, symptoms of RVFV infection mainly include a self-limiting febrile illness. However, in some cases, infected individuals can also experience hemorrhagic fever, neurological disorders, liver failure, and blindness, which could collectively be lethal. The ability of RVFV to expand geographically outside sub-Saharan Africa is of concern, particularly to the Americas, where native mosquito species are capable of virus transmission. Currently, there are no FDA-approved therapeutics to treat RVFV infection, and thus, there is an urgent need to understand the mechanisms by which the virus hijacks the host cell machinery to replicate. The significance of our research is in identifying the cellular target of sorafenib that inhibits RVFV propagation, so that this information can be used as a tool for the further development of therapeutics used to treat RVFV infection.
Ashwini Brahms; Rajini Mudhasani; Chelsea Pinkham; Krishna Kota; Farooq Nasar; Rouzbeh Zamani; Sina Bavari; Kylene Kehn-Hall. Sorafenib Impedes Rift Valley Fever Virus Egress by Inhibiting Valosin-Containing Protein Function in the Cellular Secretory Pathway. Journal of Virology 2017, 91, e00968-17 .
AMA StyleAshwini Brahms, Rajini Mudhasani, Chelsea Pinkham, Krishna Kota, Farooq Nasar, Rouzbeh Zamani, Sina Bavari, Kylene Kehn-Hall. Sorafenib Impedes Rift Valley Fever Virus Egress by Inhibiting Valosin-Containing Protein Function in the Cellular Secretory Pathway. Journal of Virology. 2017; 91 (21):e00968-17.
Chicago/Turabian StyleAshwini Brahms; Rajini Mudhasani; Chelsea Pinkham; Krishna Kota; Farooq Nasar; Rouzbeh Zamani; Sina Bavari; Kylene Kehn-Hall. 2017. "Sorafenib Impedes Rift Valley Fever Virus Egress by Inhibiting Valosin-Containing Protein Function in the Cellular Secretory Pathway." Journal of Virology 91, no. 21: e00968-17.
Unprotected sexual intercourse between persons residing in or traveling from regions with Zika virus transmission is a risk factor for infection. To model risk for infection after sexual intercourse, we inoculated rhesus and cynomolgus macaques with Zika virus by intravaginal or intrarectal routes. In macaques inoculated intravaginally, we detected viremia and virus RNA in 50% of macaques, followed by seroconversion. In macaques inoculated intrarectally, we detected viremia, virus RNA, or both, in 100% of both species, followed by seroconversion. The magnitude and duration of infectious virus in the blood of macaques suggest humans infected with Zika virus through sexual transmission will likely generate viremias sufficient to infect competent mosquito vectors. Our results indicate that transmission of Zika virus by sexual intercourse might serve as a virus maintenance mechanism in the absence of mosquito-to-human transmission and could increase the probability of establishment and spread of Zika virus in regions where this virus is not present.
Andrew Haddow; Aysegul Nalca; Franco D. Rossi; Lynn J. Miller; Michael Wiley; Unai Perez-Sautu; Samuel C. Washington; Sarah Norris; Suzanne Wollen-Roberts; Joshua D. Shamblin; Adrienne Kimmel; Holly A. Bloomfield; Stephanie M. Valdez; Thomas R. Sprague; Lucia M. Principe; Stephanie A. Bellanca; Stephanie S. Cinkovich; Luis Lugo-Roman; Lisa Cazares; William D. Pratt; Gustavo Palacios; Sina Bavari; Margaret Louise Pitt; Farooq Nasar. High Infection Rates for Adult Macaques after Intravaginal or Intrarectal Inoculation with Zika Virus. Emerging Infectious Diseases 2017, 23, 1274 -1281.
AMA StyleAndrew Haddow, Aysegul Nalca, Franco D. Rossi, Lynn J. Miller, Michael Wiley, Unai Perez-Sautu, Samuel C. Washington, Sarah Norris, Suzanne Wollen-Roberts, Joshua D. Shamblin, Adrienne Kimmel, Holly A. Bloomfield, Stephanie M. Valdez, Thomas R. Sprague, Lucia M. Principe, Stephanie A. Bellanca, Stephanie S. Cinkovich, Luis Lugo-Roman, Lisa Cazares, William D. Pratt, Gustavo Palacios, Sina Bavari, Margaret Louise Pitt, Farooq Nasar. High Infection Rates for Adult Macaques after Intravaginal or Intrarectal Inoculation with Zika Virus. Emerging Infectious Diseases. 2017; 23 (8):1274-1281.
Chicago/Turabian StyleAndrew Haddow; Aysegul Nalca; Franco D. Rossi; Lynn J. Miller; Michael Wiley; Unai Perez-Sautu; Samuel C. Washington; Sarah Norris; Suzanne Wollen-Roberts; Joshua D. Shamblin; Adrienne Kimmel; Holly A. Bloomfield; Stephanie M. Valdez; Thomas R. Sprague; Lucia M. Principe; Stephanie A. Bellanca; Stephanie S. Cinkovich; Luis Lugo-Roman; Lisa Cazares; William D. Pratt; Gustavo Palacios; Sina Bavari; Margaret Louise Pitt; Farooq Nasar. 2017. "High Infection Rates for Adult Macaques after Intravaginal or Intrarectal Inoculation with Zika Virus." Emerging Infectious Diseases 23, no. 8: 1274-1281.
Ebola virus (EBOV) infection results in high morbidity and mortality and is primarily transmitted in communities by contact with infectious bodily fluids. While clinical and experimental evidence indicates that EBOV is transmitted via mucosal exposure, the ability of non-biting muscid flies to mechanically transmit EBOV following exposure to the face had not been assessed. To investigate this transmission route, house flies (Musca domestica Linnaeus) were used to deliver an EBOV/blood mixture to the ocular/nasal/oral facial mucosa of four cynomolgus macaques (Macaca fascicularis Raffles). Following exposure, macaques were monitored for evidence of infection through the conclusion of the study, days 57 and 58. We found no evidence of systemic infection in any of the exposed macaques. The results of this study indicate that there is a low potential for the mechanical transmission of EBOV via house flies - the conditions in this study were not sufficient to initiate infection.
Andrew D. Haddow; Farooq Nasar; Christopher W. Schellhase; Roger D. Moon; Susana L. Padilla; Xiankun Zeng; Suzanne E. Wollen-Roberts; Joshua D. Shamblin; Elizabeth C. Grimes; Justine M. Zelko; Kenneth J. Linthicum; Sina Bavari; Margaret Louise Pitt; John C. Trefry. Low potential for mechanical transmission of Ebola virus via house flies (Musca domestica). Parasites & Vectors 2017, 10, 218 .
AMA StyleAndrew D. Haddow, Farooq Nasar, Christopher W. Schellhase, Roger D. Moon, Susana L. Padilla, Xiankun Zeng, Suzanne E. Wollen-Roberts, Joshua D. Shamblin, Elizabeth C. Grimes, Justine M. Zelko, Kenneth J. Linthicum, Sina Bavari, Margaret Louise Pitt, John C. Trefry. Low potential for mechanical transmission of Ebola virus via house flies (Musca domestica). Parasites & Vectors. 2017; 10 (1):218.
Chicago/Turabian StyleAndrew D. Haddow; Farooq Nasar; Christopher W. Schellhase; Roger D. Moon; Susana L. Padilla; Xiankun Zeng; Suzanne E. Wollen-Roberts; Joshua D. Shamblin; Elizabeth C. Grimes; Justine M. Zelko; Kenneth J. Linthicum; Sina Bavari; Margaret Louise Pitt; John C. Trefry. 2017. "Low potential for mechanical transmission of Ebola virus via house flies (Musca domestica)." Parasites & Vectors 10, no. 1: 218.
The demonstrated clinical efficacy of a recombinant vesicular stomatitis virus (rVSV) vaccine vector has stimulated the investigation of additional serologically distinct Vesiculovirus vectors as therapeutic and/or prophylactic vaccine vectors to combat emerging viral diseases. Among these viral threats are the encephalitic alphaviruses Venezuelan equine encephalitis virus (VEEV) and Eastern equine encephalitis virus (EEEV), which have demonstrated potential for natural disease outbreaks, yet no licensed vaccines are available in the event of an epidemic. Here we report the rescue of recombinant Isfahan virus (rISFV) from genomic cDNA as a potential new vaccine vector platform. The rISFV genome was modified to attenuate virulence and express the VEEV and EEEV E2/E1 surface glycoproteins as vaccine antigens. A single dose of the rISFV vaccine vectors elicited neutralizing antibody responses and protected mice from lethal VEEV and EEEV challenges at 1 month postvaccination as well as lethal VEEV challenge at 8 months postvaccination. A mixture of rISFV vectors expressing the VEEV and EEEV E2/E1 glycoproteins also provided durable, single-dose protection from lethal VEEV and EEEV challenges, demonstrating the potential for a multivalent vaccine formulation. These findings were paralleled in studies with an attenuated form of rVSV expressing the VEEV E2/E1 glycoproteins. Both the rVSV and rISFV vectors were attenuated by using an approach that has demonstrated safety in human trials of an rVSV/HIV-1 vaccine. Vaccines based on either of these vaccine vector platforms may present a safe and effective approach to prevent alphavirus-induced disease in humans. IMPORTANCE This work introduces rISFV as a novel vaccine vector platform that is serologically distinct and phylogenetically distant from VSV. The rISFV vector has been attenuated by an approach used for an rVSV vector that has demonstrated safety in clinical studies. The vaccine potential of the rISFV vector was investigated in a well-established alphavirus disease model. The findings indicate the feasibility of producing a safe, efficacious, multivalent vaccine against the encephalitic alphaviruses VEEV and EEEV, both of which can cause fatal disease. This work also demonstrates the efficacy of an attenuated rVSV vector that has already demonstrated safety and immunogenicity in multiple HIV-1 phase I clinical studies. The absence of serological cross-reactivity between rVSV and rISFV and their phylogenetic divergence within the Vesiculovirus genus indicate potential for two stand-alone vaccine vector platforms that could be used to target multiple bacterial and/or viral agents in successive immunization campaigns or as heterologous prime-boost agents.
Farooq Nasar; Demetrius Matassov; Robert L. Seymour; Theresa Latham; Rodion V. Gorchakov; Rebecca M. Nowak; Grace Leal; Stefan Hamm; John H. Eldridge; Robert B. Tesh; David K. Clarke; Scott C. Weaver. Recombinant Isfahan Virus and Vesicular Stomatitis Virus Vaccine Vectors Provide Durable, Multivalent, Single-Dose Protection against Lethal Alphavirus Challenge. Journal of Virology 2017, 91, e01729-16 .
AMA StyleFarooq Nasar, Demetrius Matassov, Robert L. Seymour, Theresa Latham, Rodion V. Gorchakov, Rebecca M. Nowak, Grace Leal, Stefan Hamm, John H. Eldridge, Robert B. Tesh, David K. Clarke, Scott C. Weaver. Recombinant Isfahan Virus and Vesicular Stomatitis Virus Vaccine Vectors Provide Durable, Multivalent, Single-Dose Protection against Lethal Alphavirus Challenge. Journal of Virology. 2017; 91 (8):e01729-16.
Chicago/Turabian StyleFarooq Nasar; Demetrius Matassov; Robert L. Seymour; Theresa Latham; Rodion V. Gorchakov; Rebecca M. Nowak; Grace Leal; Stefan Hamm; John H. Eldridge; Robert B. Tesh; David K. Clarke; Scott C. Weaver. 2017. "Recombinant Isfahan Virus and Vesicular Stomatitis Virus Vaccine Vectors Provide Durable, Multivalent, Single-Dose Protection against Lethal Alphavirus Challenge." Journal of Virology 91, no. 8: e01729-16.
New vaccine approaches that safely elicit immunity are needed to protect against infectious disease. Erasmus et al. report their development of an insect-virus-based platform that they use to engineer a protective vaccine against chikungunya fever. Traditionally, vaccine development involves tradeoffs between immunogenicity and safety. Live-attenuated vaccines typically offer rapid and durable immunity but have reduced safety when compared to inactivated vaccines. In contrast, the inability of inactivated vaccines to replicate enhances safety at the expense of immunogenicity, often necessitating multiple doses and boosters. To overcome these tradeoffs, we developed the insect-specific alphavirus, Eilat virus (EILV), as a vaccine platform. To address the chikungunya fever (CHIKF) pandemic, we used an EILV cDNA clone to design a chimeric virus containing the chikungunya virus (CHIKV) structural proteins. The recombinant EILV/CHIKV was structurally identical at 10 Å to wild-type CHIKV, as determined by single-particle cryo-electron microscopy, and it mimicked the early stages of CHIKV replication in vertebrate cells from attachment and entry to viral RNA delivery. Yet the recombinant virus remained completely defective for productive replication, providing a high degree of safety. A single dose of EILV/CHIKV produced in mosquito cells elicited rapid (within 4 d) and long-lasting (>290 d) neutralizing antibodies that provided complete protection in two different mouse models. In nonhuman primates, EILV/CHIKV elicited rapid and robust immunity that protected against viremia and telemetrically monitored fever. Our EILV platform represents the first structurally native application of an insect-specific virus in preclinical vaccine development and highlights the potential application of such viruses in vaccinology.
Jesse H. Erasmus; Albert Auguste; Jason T. Kaelber; Huanle Luo; Shannan L. Rossi; Karla Fenton; Grace Leal; Dal Young Kim; Jason T Kaelber Wah Chiu; Tian Wang; Ilya Frolov; Farooq Nasar; Jesse H Erasmus Albert J Auguste Huanle Luo Shannan L Rossi Karla Fenton Grace Leal Tian Wang Scott C Weaver. A chikungunya fever vaccine utilizing an insect-specific virus platform. Nature Medicine 2016, 23, 192 -199.
AMA StyleJesse H. Erasmus, Albert Auguste, Jason T. Kaelber, Huanle Luo, Shannan L. Rossi, Karla Fenton, Grace Leal, Dal Young Kim, Jason T Kaelber Wah Chiu, Tian Wang, Ilya Frolov, Farooq Nasar, Jesse H Erasmus Albert J Auguste Huanle Luo Shannan L Rossi Karla Fenton Grace Leal Tian Wang Scott C Weaver. A chikungunya fever vaccine utilizing an insect-specific virus platform. Nature Medicine. 2016; 23 (2):192-199.
Chicago/Turabian StyleJesse H. Erasmus; Albert Auguste; Jason T. Kaelber; Huanle Luo; Shannan L. Rossi; Karla Fenton; Grace Leal; Dal Young Kim; Jason T Kaelber Wah Chiu; Tian Wang; Ilya Frolov; Farooq Nasar; Jesse H Erasmus Albert J Auguste Huanle Luo Shannan L Rossi Karla Fenton Grace Leal Tian Wang Scott C Weaver. 2016. "A chikungunya fever vaccine utilizing an insect-specific virus platform." Nature Medicine 23, no. 2: 192-199.
Zika virus (ZIKV) has extended its known geographic distribution to the New World and is now responsible for severe clinical complications in a subset of patients. While substantial genetic and vector susceptibility data exist for ZIKV, less is known for the closest related flavivirus, Spondweni virus (SPONV). Both ZIKV and SPONV have been known to circulate in Africa since the mid-1900s, but neither has been genetically characterized by gene and compared in parallel. Furthermore, the susceptibility of peridomestic mosquito species incriminated or suspected in the transmission of ZIKV to SPONV was unknown. In this study, two geographically distinct strains of SPONV were genetically characterized and compared to nine genetically and geographically distinct ZIKV strains. Additionally, the susceptibility of both SPONV strains was determined in three mosquito species. The open reading frame (ORF) of the SPONV 1952 Nigerian Chuku strain, exhibited a nucleotide and amino acid identity of 97.8% and 99.2%, respectively, when compared to the SPONV 1954 prototype South African SA Ar 94 strain. The ORF of the SPONV Chuku strain exhibited a nucleotide and amino acid identity that ranged from 68.3% to 69.0% and 74.6% to 75.0%, respectively, when compared to nine geographically and genetically distinct strains of ZIKV. The ORF of the nine African and Asian lineage ZIKV strains exhibited limited nucleotide divergence. Aedes aegypti, Ae. albopictus and Culex quinquefasciatus susceptibility and dissemination was low or non-existent following artificial infectious blood feeding of moderate doses of both SPONV strains. SPONV and ZIKV nucleotide and amino acid divergence coupled with differences in geographic distribution, ecology and vector species support previous reports that these viruses are separate species. Furthermore, the low degree of SPONV infection or dissemination in Ae. albopictus, Ae. aegypti and Cx. quinquefasciatus following exposure to two geographically and genetically distinct virus strains suggest a low potential for these species to serve as vectors. Spondweni virus (SPONV) is a mosquito-transmitted flavivirus reported in Africa. Human infection with SPONV may result in a febrile illness similar to symptomatic Zika virus (ZIKV) infection, as well as many other tropical infections. Previously, little was known about the genetic relationships between SPONV and ZIKV. Additionally, the ability of SPONV to infect peridomestic mosquito species suspected or incriminated in the transmission of ZIKV was unknown. Both SPONV strains exhibited a high degree of nucleotide and amino acid identity to each other, but considerable nucleotide and amino acid divergence to ZIKV. The open reading frame (ORF) of the nine African and Asian lineage ZIKV strains originally isolated in West Africa, Central Africa, East Africa, Southeast Asia, the Pacific Islands and the New World all exhibited limited nucleotide divergence. Both strains of SPONV exhibited a low degree of infection and/or dissemination in Aedes albopictus, Ae. aegypti and Culex quinquefasciatus mosquitoes suggesting that these species have a low potential to serve as vectors. These results coupled with differences in geographic distribution, ecology and vector species indicate that SPONV and ZIKV are similar but separate species.
Andrew D. Haddow; Farooq Nasar; Hilda Guzman; Alongkot Ponlawat; Richard G. Jarman; Robert B. Tesh; Scott C. Weaver. Genetic Characterization of Spondweni and Zika Viruses and Susceptibility of Geographically Distinct Strains of Aedes aegypti, Aedes albopictus and Culex quinquefasciatus (Diptera: Culicidae) to Spondweni Virus. PLOS Neglected Tropical Diseases 2016, 10, e0005083 .
AMA StyleAndrew D. Haddow, Farooq Nasar, Hilda Guzman, Alongkot Ponlawat, Richard G. Jarman, Robert B. Tesh, Scott C. Weaver. Genetic Characterization of Spondweni and Zika Viruses and Susceptibility of Geographically Distinct Strains of Aedes aegypti, Aedes albopictus and Culex quinquefasciatus (Diptera: Culicidae) to Spondweni Virus. PLOS Neglected Tropical Diseases. 2016; 10 (10):e0005083.
Chicago/Turabian StyleAndrew D. Haddow; Farooq Nasar; Hilda Guzman; Alongkot Ponlawat; Richard G. Jarman; Robert B. Tesh; Scott C. Weaver. 2016. "Genetic Characterization of Spondweni and Zika Viruses and Susceptibility of Geographically Distinct Strains of Aedes aegypti, Aedes albopictus and Culex quinquefasciatus (Diptera: Culicidae) to Spondweni Virus." PLOS Neglected Tropical Diseases 10, no. 10: e0005083.
Diverse pathogenic agents often utilize overlapping host networks, and hub proteins within these networks represent attractive targets for broad-spectrum drugs. Using bacterial toxins, we describe a new approach for discovering broad-spectrum therapies capable of inhibiting host proteins that mediate multiple pathogenic pathways. This approach can be widely used, as it combines genetic-based target identification with cell survival-based and protein function-based multiplex drug screens, and concurrently discovers therapeutic compounds and their protein targets. Using B-lymphoblastoid cells derived from the HapMap Project cohort of persons of African, European, and Asian ancestry we identified host caspases as hub proteins that mediate the lethality of multiple pathogenic agents. We discovered that an approved drug, Bithionol, inhibits host caspases and also reduces the detrimental effects of anthrax lethal toxin, diphtheria toxin, cholera toxin, Pseudomonas aeruginosa exotoxin A, Botulinum neurotoxin, ricin, and Zika virus. Our study reveals the practicality of identifying host proteins that mediate multiple disease pathways and discovering broad-spectrum therapies that target these hub proteins.
William Leonardi; Leeor Zilbermintz; Luisa W. Cheng; Josue Zozaya; Sharon H. Tran; Jeffrey H. Elliott; Kseniya Polukhina; Robert Manasherob; Amy Li; Xiaoli Chi; Dima Gharaibeh; Tara Kenny; Rouzbeh Zamani; Veronica Soloveva; Andrew Haddow; Farooq Nasar; Sina Bavari; Michael C. Bassik; Stanley N. Cohen; Anastasia Levitin; Mikhail Martchenko. Bithionol blocks pathogenicity of bacterial toxins, ricin and Zika virus. Scientific Reports 2016, 6, srep34475 .
AMA StyleWilliam Leonardi, Leeor Zilbermintz, Luisa W. Cheng, Josue Zozaya, Sharon H. Tran, Jeffrey H. Elliott, Kseniya Polukhina, Robert Manasherob, Amy Li, Xiaoli Chi, Dima Gharaibeh, Tara Kenny, Rouzbeh Zamani, Veronica Soloveva, Andrew Haddow, Farooq Nasar, Sina Bavari, Michael C. Bassik, Stanley N. Cohen, Anastasia Levitin, Mikhail Martchenko. Bithionol blocks pathogenicity of bacterial toxins, ricin and Zika virus. Scientific Reports. 2016; 6 (1):srep34475.
Chicago/Turabian StyleWilliam Leonardi; Leeor Zilbermintz; Luisa W. Cheng; Josue Zozaya; Sharon H. Tran; Jeffrey H. Elliott; Kseniya Polukhina; Robert Manasherob; Amy Li; Xiaoli Chi; Dima Gharaibeh; Tara Kenny; Rouzbeh Zamani; Veronica Soloveva; Andrew Haddow; Farooq Nasar; Sina Bavari; Michael C. Bassik; Stanley N. Cohen; Anastasia Levitin; Mikhail Martchenko. 2016. "Bithionol blocks pathogenicity of bacterial toxins, ricin and Zika virus." Scientific Reports 6, no. 1: srep34475.
Recent experimentation with the variants of the Ebola virus that differ in the glycoprotein’s poly-uridine site, which dictates the form of glycoprotein produced through a transcriptional stutter, has resulted in questions regarding the pathogenicity and lethality of the stocks used to develop products currently undergoing human clinical trials to combat the disease. In order to address these concerns and prevent the delay of these critical research programs, we designed an experiment that permitted us to intramuscularly challenge statistically significant numbers of naïve and vaccinated cynomolgus macaques with either a 7U or 8U variant of the Ebola virus, Kikwit isolate. In naïve animals, no difference in survivorship was observed; however, there was a significant delay in the disease course between the two groups. Significant differences were also observed in time-of-fever, serum chemistry, and hematology. In vaccinated animals, there was no statistical difference in survivorship between either challenge groups, with two succumbing in the 7U group compared to 1 in the 8U challenge group. In summary, survivorship was not affected, but the Ebola virus disease course in nonhuman primates is temporally influenced by glycoprotein poly-U editing site populations.
John C. Trefry; Suzanne E. Wollen; Farooq Nasar; Joshua D. Shamblin; Steven J. Kern; Jeremy J. Bearss; Michelle A. Jefferson; Taylor B. Chance; Jeffery R. Kugelman; Jason T. Ladner; Anna N. Honko; Dean J. Kobs; Morgan Q.S. Wending; Carol L. Sabourin; William D. Pratt; Gustavo F. Palacios; M. Louise M. Pitt. Ebola Virus Infections in Nonhuman Primates Are Temporally Influenced by Glycoprotein Poly-U Editing Site Populations in the Exposure Material. Viruses 2015, 7, 6739 -6754.
AMA StyleJohn C. Trefry, Suzanne E. Wollen, Farooq Nasar, Joshua D. Shamblin, Steven J. Kern, Jeremy J. Bearss, Michelle A. Jefferson, Taylor B. Chance, Jeffery R. Kugelman, Jason T. Ladner, Anna N. Honko, Dean J. Kobs, Morgan Q.S. Wending, Carol L. Sabourin, William D. Pratt, Gustavo F. Palacios, M. Louise M. Pitt. Ebola Virus Infections in Nonhuman Primates Are Temporally Influenced by Glycoprotein Poly-U Editing Site Populations in the Exposure Material. Viruses. 2015; 7 (12):6739-6754.
Chicago/Turabian StyleJohn C. Trefry; Suzanne E. Wollen; Farooq Nasar; Joshua D. Shamblin; Steven J. Kern; Jeremy J. Bearss; Michelle A. Jefferson; Taylor B. Chance; Jeffery R. Kugelman; Jason T. Ladner; Anna N. Honko; Dean J. Kobs; Morgan Q.S. Wending; Carol L. Sabourin; William D. Pratt; Gustavo F. Palacios; M. Louise M. Pitt. 2015. "Ebola Virus Infections in Nonhuman Primates Are Temporally Influenced by Glycoprotein Poly-U Editing Site Populations in the Exposure Material." Viruses 7, no. 12: 6739-6754.
In December of 2013, chikungunya virus (CHIKV), an alphavirus in the family Togaviridae, was introduced to the island of Saint Martin in the Caribbean, resulting in the first autochthonous cases reported in the Americas. As of January 2015, local and imported CHIKV has been reported in 50 American countries with over 1.1 million suspected cases. CHIKV causes a severe arthralgic disease for which there are no approved vaccines or therapeutics. Furthermore, the lack of a commercially available, sensitive, and affordable diagnostic assay limits surveillance and control efforts. To address this issue, we utilized an insect-specific alphavirus, Eilat virus (EILV), to develop a diagnostic antigen that does not require biosafety containment facilities to produce. We demonstrated that EILV/CHIKV replicates to high titers in insect cells and can be applied directly in enzyme-linked immunosorbent assays without inactivation, resulting in highly sensitive detection of recent and past CHIKV infection, and outperforming traditional antigen preparations. We have developed an innovative approach to production of alphavirus antigens for use in diagnostic assays that results in reduced production complexity as well as improved sensitivity in application. By generating recombinant viruses that contain the structural protein genes of pathogenic alphaviruses and the nonstructural protein genes of an insect-specific alphavirus, Eilat virus, we have been able to produce insect-restricted viruses that are antigenically identical to their pathogenic counterparts. The insect-specific nature of these chimeric viruses yields an advantageous safety profile and allows for safe handling of the antigen at the bench top. Traditional antigens, produced from wild-type virus, require extensive processing, from growth at biosafety level 3 to concentration and inactivation, followed by lyophilization, which often compromises antigen reactivity and is financially costly. Furthermore, current inactivation methods are imperfect processes that have historically resulted in residual live virus and subsequent breach of containment when used in a diagnostic setting. Other approaches, such as recombinant antigens generated from viral particle subunits, are missing conformational epitopes and their application results in reduced sensitivity. Here we describe the development of a diagnostic assay using this technology for the detection of chikungunya infection in humans.
Jesse H. Erasmus; James Needham; Syamal Raychaudhuri; Michael S. Diamond; David W. C. Beasley; Stan Morkowski; Henrik Salje; Ildefonso Fernandez Salas; Dal Young Kim; Ilya Frolov; Farooq Nasar; Scott C. Weaver. Utilization of an Eilat Virus-Based Chimera for Serological Detection of Chikungunya Infection. PLOS Neglected Tropical Diseases 2015, 9, e0004119 .
AMA StyleJesse H. Erasmus, James Needham, Syamal Raychaudhuri, Michael S. Diamond, David W. C. Beasley, Stan Morkowski, Henrik Salje, Ildefonso Fernandez Salas, Dal Young Kim, Ilya Frolov, Farooq Nasar, Scott C. Weaver. Utilization of an Eilat Virus-Based Chimera for Serological Detection of Chikungunya Infection. PLOS Neglected Tropical Diseases. 2015; 9 (10):e0004119.
Chicago/Turabian StyleJesse H. Erasmus; James Needham; Syamal Raychaudhuri; Michael S. Diamond; David W. C. Beasley; Stan Morkowski; Henrik Salje; Ildefonso Fernandez Salas; Dal Young Kim; Ilya Frolov; Farooq Nasar; Scott C. Weaver. 2015. "Utilization of an Eilat Virus-Based Chimera for Serological Detection of Chikungunya Infection." PLOS Neglected Tropical Diseases 9, no. 10: e0004119.